Na busca de alternativas para mensurar a emissão foliar de amônia (NH3) e minimizar as perdas de N-NH3 em pastagens, foram realizados três trabalhos. Os dois primeiros com objetivo de verificar se o absorvedor com espuma, já utilizado na quantificação da volatilização de N-NH3, também é eficiente para mensurar a emissão foliar, sem causar alterações no processo de perda de nitrogênio. O terceiro trabalho, realizado em pastagem de capim Colonião (Panicum maximum Jacq. cv. Colonião) no verão, inverno e primavera, avaliou o efeito da aplicação de lâminas de água, após a adubação com uréia, sobre as perdas de N-NH3 do solo por volatilização e emissão foliar. O absorvedor de amônia com espuma não causa alteração no processo de perda de N-NH3 e colocado 1 cm acima das folhas superiores, é efetivo em capturar o N-NH3 perdido por emissão foliar da pastagem, quando se fertiliza em superfície com nitrato de amônio e uréia. A aplicação de água imediatamente após a adubação com uréia é eficiente para reduzir as perdas de NNH3 por volatilização. No verão, a aplicação de 3,2 mm de água foi suficiente para reduzir as perdas de N-NH3 para menos de 3,1 % do N aplicado, enquanto na ausência de irrigação ocorreram perdas de 30,5%. A taxa de volatilização é influenciada pela quantidade de água disponível no solo, sendo baixa quando a uréia é aplicada em solo seco ou quando o solo seca rapidamente, mesmo que a temperatura ambiente seja elevada. A emissão foliar de N-NH3 não foi influenciada pela aplicação ou não de água, após a adubação com uréia. / In search of alternatives to measure ammonia (NH3) foliar emission and minimize N-NH3 losses in pasture three research works were accomplished. The two first works aimed at checking whether or not the foam absorber, which was already used to quantify N-NH3 volatilization, is also efficient to measure foliar emission without interfering in nitrogen loss process. The third one was performed in Panicum maximum Jacq. cv. Colonião pasture during three different seasons and evaluated the use of irrigation levels after urea fertilization on N-NH3 losses through volatilization and foliar emission. The ammonia foam absorber does not alter N-NH3 loss process and when place at height of 1 cm from the upper leaves it is effective in capturing N-NH3 lost through foliar emission when fertilization is done superficially with ammonium nitrate and urea. Water application immediately after fertilization is efficient to reduce N-NH3 losses through volatilization. During summer the use of 3.2 mm water was enough to decrease N-NH3 loss to less than 3.1% of applied N, while the lack of irrigation caused 30.5% losses. Volatilization rate is influenced by the quantity of water available in the soil, being low when urea is applied to dry soil or when the soil dries fast even if the environment temperature is high. N-NH3 foliar emission was not influenced by water application after urea fertilization.

O presente trabalho foi desenvolvido no Departamento de Zootecnia da Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de São Paulo FZEA/USP localizada em Pirassununga-SP no período de 16/12/2006 a 10/05/2007 em condições de campo com o cultivo do híbrido do milho super precoce GNZ-2005 em Latossolo Vermelho Amarelo distrófico férrico. O experimento teve como objetivo avaliar a eficiência de fertilizante nitrogenado FASN (Fusion Amonium Sulfate Nitrate) comparado na composição químico-bromatológica e algumas características agronomicas da cultura de milho (Zea mays L.) à fontes tradicionais de nitrogênio como nitrato de amônio, uréia e sulfato de amônio, implantado no sistema plantio direto. Foram avaliadas as perdas de nitrogênio por volatilização da amônia, os atributos químicos e físicos do solo, rendimento de grãos (kg ha-1), matéria seca total (%) e análise bromatológica e foliar. A dose de nitrogênio foi 100 kg ha-1 de N em todos os tratamentos, exceto nas testemunhas (T1 e T2), que também receberam 450 kg ha-1 de 8:28:16 + 0,5% de Zn. O delineamento experimental foi em blocos completos casualizados, com dez tratamentos e quatro repetições. Os tratamentos foram: T1: Testemunha +Gesso (-N+S), T2: Testemunha absoluta (-N-S), T3: Uréia - 45% N, T4: Uréia + Gesso 14%S, T5: Uréia + sulfato de amônio - 21%N /24%S-(33:00:00+12%S), T6: Nitrato de amônio - 30%N, T7: Nitrato de amônio - 30%N + gesso, T8: Nitrato de amônio + Sulfato de Amônio - 21% N/ 24%S - (27,8:00:00+6%S), T9: Sulfato de amônio, T10: FASN 26%N/14%S. Perdas por volatilização de amônia foram avaliadas em todos os tratamentos de T1 a T10 por meio de coletores semi-abertos estáticos colocados em cada parcela em que se estudou a volatilização. Foram colocadas cinco bases para cada coletor, onde se aplicou as fontes nitrogenadas na dose de 100 kg ha-1 de N. O N-NH3 volatilizado foi retido em espumas embebidas em solução ácida e extraída com solução ácido clorídrico para posterior quantificação das perdas. A avaliação do estado nutricional das plantas, de atributos químicos e físicos do solo, volatilização de amônia, características bromatológicas e a produtividade foram efetuadas em todos os tratamentos. A avaliação do estado nutricional foi feita pela análise química dos teores de nutrientes nas folhas coletadas no florescimento. Atributos químicos do solo foram avaliados por meio de análises laboratoriais de amostras coletadas nas camadas de 0 a 20 e de 20 a 40 cm de profundidade, após a colheita do milho. Dentre das fontes nitrogenadas avaliadas, a uréia implicou em maiores perdas de nitrogênio N-NH3 aos seis dias após aplicação. Com relação ao desempenho geral das fontes de nitrogênio, pode-se constatar que o FASN teve comportamento similar ao do sulfato de amônio e do nitrato de amônio. / O presente trabalho foi desenvolvido no Departamento de Zootecnia da Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de São Paulo FZEA/USP localizada em Pirassununga-SP no período de 16/12/2006 a 10/05/2007 em condições de campo com o cultivo do híbrido do milho super precoce GNZ-2005 em Latossolo Vermelho Amarelo distrófico férrico. O experimento teve como objetivo avaliar a eficiência de fertilizante nitrogenado FASN (Fusion Amonium Sulfate Nitrate) comparado na composição químico-bromatológica e algumas características agronomicas da cultura de milho (Zea mays L.) à fontes tradicionais de nitrogênio como nitrato de amônio, uréia e sulfato de amônio, implantado no sistema plantio direto. Foram avaliadas as perdas de nitrogênio por volatilização da amônia, os atributos químicos e físicos do solo, rendimento de grãos (kg ha-1), matéria seca total (%) e análise bromatológica e foliar. A dose de nitrogênio foi 100 kg ha-1 de N em todos os tratamentos, exceto nas testemunhas (T1 e T2), que também receberam 450 kg ha-1 de 8:28:16 + 0,5% de Zn. O delineamento experimental foi em blocos completos casualizados, com dez tratamentos e quatro repetições. Os tratamentos foram: T1: Testemunha +Gesso (-N+S), T2: Testemunha absoluta (-N-S), T3: Uréia - 45% N, T4: Uréia + Gesso 14%S, T5: Uréia + sulfato de amônio - 21%N /24%S-(33:00:00+12%S), T6: Nitrato de amônio - 30%N, T7: Nitrato de amônio - 30%N + gesso, T8: Nitrato de amônio + Sulfato de Amônio - 21% N/ 24%S - (27,8:00:00+6%S), T9: Sulfato de amônio, T10: FASN 26%N/14%S. Perdas por volatilização de amônia foram avaliadas em todos os tratamentos de T1 a T10 por meio de coletores semi-abertos estáticos colocados em cada parcela em que se estudou a volatilização. Foram colocadas cinco bases para cada coletor, onde se aplicou as fontes nitrogenadas na dose de 100 kg ha-1 de N. O N-NH3 volatilizado foi retido em espumas embebidas em solução ácida e extraída com solução ácido clorídrico para posterior quantificação das perdas. A avaliação do estado nutricional das plantas, de atributos químicos e físicos do solo, volatilização de amônia, características bromatológicas e a produtividade foram efetuadas em todos os tratamentos. A avaliação do estado nutricional foi feita pela análise química dos teores de nutrientes nas folhas coletadas no florescimento. Atributos químicos do solo foram avaliados por meio de análises laboratoriais de amostras coletadas nas camadas de 0 a 20 e de 20 a 40 cm de profundidade, após a colheita do milho. Dentre das fontes nitrogenadas avaliadas, a uréia implicou em maiores perdas de nitrogênio N-NH3 aos seis dias após aplicação. Com relação ao desempenho geral das fontes de nitrogênio, pode-se constatar que o FASN teve comportamento similar ao do sulfato de amônio e do nitrato de amônio.

Amostras da água subterrânea foram coletadas em poços piezométricos localizados em dois transectos ao longo das vertentes e dois transectos ao longo da cabeceira de drenagem de uma microbacia experimental, durante o período de maio de 2005 a abril de 2006. Em cada transecto os poços piezométricos estavam localizados respectivamente na parte alta da vertente, sob floresta plantada de Eucalyptus, na meia encosta, sob mata ciliar, e adjacente ao riacho. As análises destas amostras também foram comparadas com a análise simultânea da água do riacho, coletada na calha da estação linimétrica da microbacia. As análises químicas incluíram principalmente nitrato (N-NO3 -) e amônio (N-NH4 +), além do ferro, silício, alumínio, carbono orgânico dissolvido (DOC) e pH. O objetivo foi inferir sobre aspectos do funcionamento hidrológico da zona ripária, em termos de alteração da concentração dos íons analisados entre os pontos amostrados. A fim de inferir sobre o efeito da vegetação e das características do solo nas modificações da água subterrânea ao longo da vertente, foi realizado inventário para determinação da área basal da vegetação e análise granulométrica e química do solo. Os principais resultados mostram que na zona ripária adjacente ao riacho há maior concentração de matéria orgânica no solo e de amônio na água subterrânea. As concentrações de amônio e nitrato na água do riacho foram sempre menores do que as concentrações nos piezômetros adjacentes ao riacho próximo ao vertedor. Isso indica que na zona ripária ocorrem modificações na água subterrânea que garantem a manutenção do ecossistema lótico. Também foi possível identificar diferenças ao longo da microbacia, entre a cabeceira de drenagem e a área próxima ao vertedor, o que demonstra que a zona ripária não é homogênea na área da microbacia. A geologia, a granulometria e o teor de matéria orgânica do solo, a precipitação, o aumento do nível freático e a influência da água subterrânea, foram os principais fatores de alteração das demais variáveis do estudo. / Groundwater samples were collected in piezometers located in two transects through the hillslope and two in the headwater of an experimental catchment from May 2005 to April 2006. In each transect there were one piezometer in the upland under the Eucalyptus planted forest, one piezometer at the middle slope in the riparian area, and the other piezometer was located by the stream with a total of 12 piezometers. The samples collected fortnightly were analyzed and compared with the stream water collected in the stream gauging station. The main variables analyzed were nitrate (N-NO3 -) and ammonium (N-NH4 +) as well as iron, silicon, aluminum, dissolved organic carbon (DOC) and pH. The objective was to infer about riparian zone hydrology, based on the difference among chemical composition of the samples. A characterization of the experimental catchment was performed from a vegetation inventory and soil analysis in order to assess the influence of soil properties and vegetation nutritional demand on the chemistry of the solutions. Chemical analyses demonstrated that in the riparian zone close to the stream soil organic matter content was higher than in other topographic positions as well as concentrations of ammonium in the groundwater throughout the year. The concentrations of ammonium and nitrate in the groundwater beside the stream were always higher. These results indicated that in riparian zone the modifications in the groundwater contribute to the lotic ecosystem maintenance. The results also indicated that the riparian zone is heterogeneous along the catchment area and that the behavior of headwater and downstream may be different. The geology, soil granulometric and organic matter level, precipitation and groundwater level were the main factors that influenced the studied variables.

Em uma das etapas do processo de produção do dióxido de urânio, obtém-se como subproduto o fluoreto de amônio em solução. O dióxido de urânio é utilizado na confecção dos elementos que abastecem às Usinas Nucleares.Na unidade fabril da empresa Indústrias Nucleares do Brasil S. A.,esse material sintético é cristalizado em um oneroso e delicado processo de secagem, utilizando grandes equipamentos e considerável quantidade de insumos.A unidade fabril em questão é capaz de produzir anualmente pouco mais de 130 toneladasde fluoreto de amônio seco e a perspectiva atual é favorável à expansão da capacidade instalada. Atualmente, não há destinação final ou aplicação prevista para esse material seco, ou seja, o material é armazenado por tempo indeterminado, onerando ainda mais a produção do combustível nuclear com os custos de manuseio e armazenagem. Este trabalho propõe-se a estudar uma rota alternativapara a síntese da fluorita, ou fluoreto de cálcio (CaF2), a partir da solução de fluoreto de amônio obtida como subproduto do processode produção do Dióxido de Urânio, verificando qual o melhor agente precipitante e estabelecendo as melhores condições para o processo de síntese. A fluorita possui uma vasta aplicação, principalmente nos campos da química, siderurgia e cerâmica. Os melhores resultados foram obtidos utilizando-se calcita (CaCO3) como agente precipitante, mantidas as condições de processo: temperatura (Treação) =70 ºC; tempo de residência (treação) = 2 h e relação estequiométrica entre os reagentes (RE) = 1:1. A calcita foi caracterizada por ICP/OES, difratometria de raios-X (DRX) e microscopia eletrônica de varredura acoplada a espectrômetro de energia dispersiva (MEV/EDS).Os produtos foram caracterizados por DRX e MEV/EDS. / At the uranium dioxide production process, ammonium fluoride is obtainedas a byproduct. Uranium dioxide is used at the production of fuel assemblies for Nuclear Mills.At Indústrias Nucleares do Brasil S. A. site, synthetic ammonium fluoride is crystallized in a costly and delicate drying process, which spends importantamounts of supplies. The industrial site in question is able to annually produce over 130 tons ofdryammonium fluoride and the current outlook is favorable to the expansion of installed capacity. Currently, there is no final destination or application provided for this dry material, i.e.,the material is stored indefinitely, further burdening the production of nuclear fuel with the handling and storage costs. This work proposes to study an alternative route for the synthesis of fluorite, or calcium fluoride (CaF2), from ammonium fluoride solution obtained as a by-product of uranium dioxide production process, checkingthe best precipitant agent and establishing the best conditions for the synthesis process.The fluorite has a wide application, particularly in the fields of chemistry, steel and ceramic. The best results were achievedusing calcite (CaCO3) as precipitating agent, keeping the process conditions: temperature (Treação) = 70 ° C; residence time (treação) = 2 h stoichiometric ratio between the reagents (ER) = 1:1.The calcite was characterized by X-ray diffraction (XRD) and scanning electron microscopy coupled with energy dispersive spectrometer (SEM/EDS).The products were characterized by XRD and SEM/EDS.

As concentrações de carbono inorgânico e compostos sulfurados a partir das quais pode haver melhoria ou inibição do processo ANAMMOX variam muito na literatura especializada. Características como o tipo de efluente, inóculo, reator, temperatura, pH e tempo de detenção hidráulica influenciam as respostas obtidas em cada trabalho. A presente pesquisa teve como objetivo encontrar uma relação ideal entre as concentrações de bicarbonato e sulfeto utilizadas pelas bactérias responsáveis pelo processo. Para isso, na etapa 1, tentou-se enriquecer biomassa ANAMMOX em um reator contínuo de leito fixo utilizando lodo de uma lagoa aerada proveniente da ETE da indústria \"Incofap\", fabricante de ração animal no município de Araraquara (São Paulo). Posteriormente, na etapa 2, para encontrar a relação ideal entre bicarbonato e sulfeto, optou-se por utilizar reatores em batelada inoculados com lodo proveniente de um reator experimental ANAMMOX previamente enriquecido. Tais reatores foram alimentados com substrato sintético contendo a mesma concentração de nitrogênio amoniacal e nitrito (70 mgN/l), enquanto variou-se as concentrações de bicarbonato (5 - 125mgHCO3-/l) e sulfeto (0 - 32 mgS2-/l) por meio de um planejamento fatorial 2 x 2. Concluiuse, na etapa 1, que o lodo da \"Incofap\" possui grande potencial para processos de nitrificação parcial e desnitrificação heterotrófica, desde que condições de microaeração e estrita anaerobiose, respectivamente, sejam satisfeitas. Enquanto isso, na etapa 2, concluiu-se que qualquer presença de sulfeto no meio prejudica o processo ANAMMOX e quanto mais próximo da razão teórica estiver a concentração de bicarbonato, melhor é a atividade microbiana. / The concentrations of inorganic carbon and sulfur compounds from which there may be improvement or inhibition of the ANAMMOX process vary greatly in the literature. Characteristics such as the type of effluent, inoculum, reactor, temperature, pH and time of hydraulic detention influence the responses obtained in each work. The present research had as objective to find an ideal relation between the concentrations of bicarbonate and sulfide used by the bacteria responsible for the process. For this, in step 1, we tried to enrich ANAMMOX biomass in a continuous bed reactor using sludge from an aerated lagoon from the WWTP of the industry \"Incofap\", animal feed manufacturer in the city of Araraquara (São Paulo). Later, in step 2, to find the ideal relationship between bicarbonate and sulfide, we decided to use batch reactors inoculated with sludge from a previously enriched ANAMMOX experimental reactor. These reactors were fed with synthetic substract containing the same concentration of ammonium nitrogen and nitrite (70 mgN/l), while varying the concentrations of bicarbonate (5 - 125 mgHCO3-/l) and sulfide (0 - 32 mgS2-/l ) by means of a 2 x 2 factorial design. In stage 1, it was concluded that the \"Incofap\" sludge has great potential for partial nitrification and heterotrophic denitrification, provided that microaeration conditions and strict anaerobiosis are respectively satisfied. Meanwhile, in step 2, it was concluded that any presence of sulfide in the medium impairs the ANAMMOX process and the closer to the theoretical ratio is the bicarbonate concentration, better is the microbial activity.

The Deepwater Horizon (DWH) oil spill in the Gulf of Mexico (GoM) in the spring of 2010 introduced 4.4 million of barrels of oil to the ecosystem. Some biodegradation of the oil occurs when microorganisms, particularly bacteria, metabolize the oil as a carbon source. During this process, the microbes also require nutrients for energy. An introduction of oil at this magnitude has the ability to induce large blooms of microbes, which in turn can affect nutrient concentrations. Microbial petroleum degradation decreases nutrient concentrations, whereas the microbial assimilation and decay of organic matter increase nutrient concentrations. This study assessed whether any changes in nitrate, ammonium, and/or phosphate concentrations from historical levels could be attributed to the oil spill as a result of biodegradation, and how those changes can impact the GoM ecosystem. Nutrient samples were collected at discrete depths throughout the water column, in a cross-shelf transect inland from the spill site outside the DeSoto Canyon, in August 2010, February 2011, and May 2011 (four months to one year after the spill). In August 2010, a subsurface oil plume was found at depths of 1000 m to 1200 m. At the same depth of ~1000 m, a significant decrease in nitrate was observed, indicating the biodegradation of oil by heterotrophic bacteria of the aphotic zone, compared to earlier data during August 2000, when no known oils were present. Overall temporal increases in ammonium and dissolved organic nitrogen (DON) were observed both in near-surface waters and at an intermediate depth of ~400 m next to the walls of the DeSoto Canyon, suggesting an incremental die-off of both plankton and benthic organisms during accelerated recycling of nutrients. Continued decreases of phosphate were observed into February 2011, supporting ongoing biodegradation then as well. By May 2011, however, there were more increases in near-surface ammonium concentrations, compared to April 2000, with the implication that continued interseasonal recycled nitrogen accumulations may have been due to a decadal ecological regime shift, after a combination of top-down overfishing, petroleum perturbations, and/or increases of toxic harmful algal blooms (HABs).

Submitted in fulfillment of the degree of Master of Applied Science: Biotechnology, Durban University of Technology, Durban, South Africa, 2017. / Domestic wastewater contains a high nutrient load, primarily in the form of Carbon (C), Nitrogen (N), and Phosphorous (P) compounds. If left untreated, these nutrients can cause eutrophication in receiving environments. Biological wastewater treatment utilizes a suspension of microorganisms that metabolize this excess nutrient load. Nitrogen removal in these systems are due to the synergistic processes of nitrification and denitrification, each of which requires its own set of operating parameters and controlling microbial groups. An alternative N-removal pathway termed the anammox process allows for total N-removal in a single step under anoxic conditions. This process, mediated by the anammox bacterial group, requires no organic carbon, produces negligible greenhouse gases and requires almost 50 % less energy than the conventional process, making it a promising new technology for efficient and cost-effective N-removal. In this study, a sequencing batch reactor (SBR) was established for the autotrophic removal of N-rich wastewater through an anammox-centric bacterial consortia. The key microbial members of this consortia were characterized and quantified over time using molecular methods and next generation sequencing to determine if the operational conditions had any effect on the seed inoculum population composition. Additionally, local South African wastewater treatment plants were screened for the presence of anammox bacteria through 16S rRNA amplification and enrichment in different reactor types. A 3 L bench scale SBR was inoculated with active biomass (~ 5 % (v/v)) sourced from a parent anammox enrichment reactor, and maintained at a temperature of 35 °C ± 1 °C. The reactor was fed with a synthetic wastewater medium containing no organic C, minimal dissolved oxygen (< 0.5 mg/L), and N in the form of ammonium and nitrite in the ratio of 1:1.3. The reactor was operated for a period of 366 days and the effluent ammonium, nitrite and nitrate were measured during this period. The hydraulic retention time was controlled at 4.55 days from Day 1 to Day 250, and thereafter shortened to 1.52 days from Day 251 to Day 360 due to an increased nitrogen removal rate (NRR). During Phase I of operation (Day 1 to Day 150), the reactor performance gradually increased up to an NRR of ~160 mg N/day. During Phase II (Day 151 to Day 250), the overall reactor performance decreased with the NRR decreasing to ~90 mg N/day, while Phase III (Day 251 to Day 366) displayed a gradual recovery of NRR back to the reactor optimum of ~160 mg N/day. The accumulation of nitrate in the effluent during the latter parts of Phase II and Phase III, coupled with oxygen ingress (~2.1 mg/L) in the same period, indicated that it was not the anammox pathway that was dominating N-removal within the reactor, but more likely the second half of the nitrification pathway mediated by the nitrite oxidizing bacteria (NOB). This was further confirmed through molecular analysis, which indicated that the bacterial population had shifted significantly over the course of reactor operation. Quantitative PCR methods displayed a decrease in all the key N-removing population groups from Day 1 to Day 140, and a marginal increase in anammox and aerobic ammonia oxidizing bacteria from Day 140 – Day 260. From Day 300 onwards, NOB had started dominating the system, simultaneously suppressing the growth of other N-removing bacterial groups. Despite this, the NRR peaked during this period, indicating an alternative mechanism for ammonia removal within the reactor system. A total population analysis using NGS was also performed, which corroborated the QPCR results and displayed a population shift away from anammox bacteria towards predominantly NOB and members of the phylum Chloroflexi. The proliferation of aerobic NOB and Chloroflexi, and the suppression of anammox bacteria, indicated that DO ingress was indeed the primary cause of the population shift within the reactor. Despite this population shift, N-removal within the reactor remained high. New pathways have recently emerged which implicate these two groups as potential N oxidizers, with specific NOB groups showing the ability for oxidation of ammonia through the comammox process, and members of the Phylum Chloroflexi being capable of nitrite reduction. This could imply that an alternate pathway was responsible for the majority of N-removal within the system, in addition to the anammox and conventional nitrification pathways. Additionally, in an attempt to detect a local anammox reservoir, eleven wastewater systems from around South Africa were screened for the presence of anammox bacteria. Through direct and nested PCR-based screening, anammox bacteria was not detectable in any of the activated sludge samples tested. Based on the operating conditions of the source wastewater systems, a subset of three sludge samples were selected for further enrichment. After 60-110 days of enrichment in multiple reactor configurations, only one reactor sample tested positive for the presence of anammox bacteria. Although this result indicates that anammox bacteria might not be ubiquitous within every biological wastewater system, it is more likely that anammox bacteria might only be present at undetectable levels, and that an extended enrichment prior to screening is necessary for a true representation of anammox bacterial prevalence in an environmental sample. / M

Les protéines de la famille Mep/Amt/Rh sont largement conservées dans l’évolution. Cette famille comprend les facteurs Rhésus dont les antigènes Rh humains sont les membres les plus notoires. Le rôle des protéines de type Mep/Amt/Rh en tant que transporteurs d’ammonium a largement été décrit chez les bactéries, les champignons et les plantes. Néanmoins, leur mécanisme de fonctionnement demeure élusif et la régulation de leur activité a été peu abordée chez les organismes eucaryotes. En utilisant comme modèles de la famille Mep/Amt/Rh les trois transporteurs d’ammonium de la levure Saccharomyces cerevisiae, nous avons tenté de comprendre les mécanismes de fonctionnement et de régulation de cette famille de protéines membranaires.<p>Nous montrons qu’un résidu aspartate, conservé dans la famille Mep/Amt/Rh et situé à proximité d’un vestibule cation-attractif, joue un rôle structural dans la reconnaissance de l’ammonium chez le transporteur Mep2. De plus, un résidu histidine très conservé dans le pore hydrophobe des protéines Mep/Amt/Rh est substitué par un aspartate chez un sous-groupe de transporteurs d’ammonium fongiques. Cette substitution permet de définir deux sous-familles fonctionnelles possédant des propriétés bien distinctes.<p>Nous montrons également que la kinase Npr1 intervient dans la modulation de l’activité intrinsèque des trois protéines Mep qui demeurent inactives mais stables à la membrane plasmique en absence de la kinase. <p>Hormis leur rôle dans le transport d’ammonium en tant que source d’azote, nous montrons que l’activité des protéines Mep est requise pour différentes réponses physiologiques. Une diminution d’entrée d’ammonium en absence des protéines Mep ou de leur régulateur positif Npr1 entraîne une dérépression des gènes soumis à la répression catabolique azotée ainsi qu’un défaut dans le repompage de l’ammonium catabolique excrété durant la croissance en présence d’autres sources azotées. Un rôle supplémentaire de senseur d’ammonium avait été attribué au transporteur Mep2 dans l’induction de la croissance filamenteuse en réponse à une limitation en ammonium. Nous montrons que l’état d’activité de la protéine Mep2 est étroitement lié à sa capacité à induire le développement filamenteux. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

De nombreuses substances chimiques sont susceptibles de mener à des phénomènes d'incompatibilité, lors de leur mise en contact avec d'autres produits ou matériaux. Pour maîtriser ces risques, une identification rapide et précise de ces incompatibilités est nécessaire. Elle est réalisée, jusqu’à présent, par des outils expérimentaux qui malgré leur incontestable importance, ne fournissent que des informations limitées. Pour les compléter et améliorer la compréhension des processus d'incompatibilité, ce travail de thèse, vise à étudier la réactivité du nitrate d'ammonium, produit très utilisé dans l'industrie, connu pour sa longue liste d'incompatibilités et impliqué dans des nombreux accidents majeurs (Toulouse 2001), à l'aide de la modélisation moléculaire. Une étude théorique approfondie, basée sur des calculs DFT, destinée à identifier les chemins réactionnels, les produits formés ainsi que la chaleur dégagée par les réactions a été menée.Après avoir caractérisé le mécanisme radicalaire de décomposition du nitrate d’ammonium pur en phase gaz, une étude détaillée de la réactivité du mélange du nitrate d’ammonium et du dichloroisocyanurate de sodium (DCCNa), a été réalisée. Un chemin réactionnel a été caractérisé, il s'agit de la réaction directe entre le nitrate d'ammonium et le DCCNa, en présence d'une molécule d'eau. Ensuite, ces méthodes théoriques ont été appliquées à d'autres systèmes d'incompatibilités chimiques avec le nitrate d'ammonium (NaNO2, substances chlorées) en focalisant sur les étapes limitantes des mécanismes afin de tester leur pertinence et potentiel en tant qu'outil de prédiction a priori du phénomène d'incompatibilité. Ce travail, pionnier dans l'étude microscopique des incompatibilités chimiques du nitrate d'ammonium, a permis de clarifier les mécanismes réactionnels lors de la décomposition du produit pur et en contact avec des contaminants. / Numerous chemical reactants tend to lead to undesired phenomena of incompatibility. In order to prevent the happening of those phenomena, a rapid and accurate identification of the incompatibilities is needed. Until now, experimental studies, which provide insightful, but limited information, were the only possible approach used for the study of incompatibilities. In this work a molecular modeling approach was used in order to complete and improve experimental results of incompatibility processes for the reactivity of ammonium nitrate. Ammonium nitrate is a widely used compound in the chemical industry, known for its long list of incompatibilities and often involved in major accidents (Toulouse, 2001).This theoretical work, based on Density Functional Theory (DFT) calculations, is intended to provide insights into the possible reaction pathways, enthalpies and products formed by incompatibility reaction between ammonium nitrate and other compounds. In this thesis the mechanism of the decomposition of pure ammonium nitrate in the gas phase was firstly characterized and then the reactivity of the mixture of ammonium nitrate – sodium dichloroisocyanurate (DCCNa) was considered. The results obtained suggest the existence of a reaction path, involving a direct reaction between the ammonium nitrate and DCCNa in the presence of a molecule of water.Furthermore, the theoretical approach was employed focusing on the study of the limiting steps of other chemical incompatibilities for ammonium nitrate systems (NaNO2, chlorinated substances). Those results were employed to understand the potentiality of the theoretical approach as a priori predicting tool for the incompatibility processes.In summary this work, pioneer in the microscopic study of chemical incompatibilities of ammonium nitrate, has clarified the reaction mechanisms involved in the decomposition of pure compounds, as well as in contact with other contaminants.

Allium sativum and Tulbaghia violacea are some of the most important medicinal plants used by South African traditional healers for the treatment of flu, fever, cold, tuberculosis, asthma and many more diseases. However, growth, yield and quality are constrained by excessive and under fertilization. This study was carried out to determine, firstly, the effect of N source (ammonium sulphate and calcium nitrate) on yield and quality of A. sativum and T. violacea plants. Secondly, to determine the best season for harvesting T. violacea and lastly, to determine the antifungal effects of A. sativum and T. violacea plant extracts against plant pathogens Altenaria solani and Sclerotium rolfsii. Both plants were treated with both N sources applied as topdressing treatments at a total of 0, 50, 100, 150 and 200 kg.ha-1, divided into three applications at three week (A. sativum) and three month (T. violacea) intervals. A. sativum plants were sampled at 54, 82, 112, 140 and 175 days after planting (DAP) while, T. violacea plants were sampled monthly for ten months. Parameters recorded were growth analysis, yield and bioactivity for both plant species. Both nitrogen sources improved plant growth and yield of A. sativum and T. violacea plants. Calcium nitrate at 150 kg•ha-1 and ammonium sulphate at 200 kg•ha-1 produced the highest at 24 t•ha-1 and 27 t•ha-1, respectively. Ammonium sulphate improved bioactivity of leaves with the highest bioactivity recorded at 82 and 112 DAP. Yield obtained from the autumn harvest was not affected by N source. Ammonium sulphate and calcium nitrate at 200 kg•ha-1 produced the highest yields of 23.6 t•ha-1 and 23.5 t•ha-1, respectively. In contrast, yield obtained from the winter harvest was affected by N source at 200 kg•ha-1, with significantly better yield of 30.8 t•ha-1 with calcium nitrate compared to 27.4 t•ha-1 with ammonium sulphate. Crude extracts of T. violacea bulbs that were treated with ammonium sulphate significantly inhibited the growth of plant pathogenic fungi, whereas extracts from plants treated with calcium nitrate showed low bioactivity. Extracts from plants grown with ammonium sulphate at 100 kg•ha-1 were more effective in controlling growth of plant pathogens when compared to other N levels. The minimum inhibitory concentration (MIC) effects of A. sativum against S. rolfsii and A. solani were at 0.01 mg•mL-1. The MIC of T. violacea extracts against A. solani was at 0.006 mg•mL<Sup>-1. The MIC of T. violacea extracts were better than previously reported in literature. Therefore, A. sativum and T. violacea plant extracts can be used as fungicides against S. rolfsii and A. solani diseases for crops such as tomato and potato. / Dissertation (MInstAgrar)--University of Pretoria, 2010. / Plant Production and Soil Science / unrestricted